SERUM BIOCHEMICAL DETERMINATIONS OF PROPOLIS-TREATED RATS

SERUM BIOCHEMICAL DETERMINATIONS OF PROPOLIS-TREATED RATS.

J.M. SFORCIN , S.R.C. FUNARI, E.L.B. NOVELLI .

1 Department of Support, Production and Animal Health - Biology - FOA - UNESP, Campus of Araçatuba, State of São Paulo, Brazil, 2 Department of Production and Animal Exploration - FMVZ - UNESP, Campus of Botucatu, State of São Paulo, Brazil, 3 Center for the Study of Venoms and Venomous Animals - CEVAP - UNESP, State of São Paulo, Brazil, 4 Department of Chemistry - IB - UNESP, Campus of Botucatu, State of São Paulo, Brazil.

ABSTRACT. Ethanolic extracts of the bee glue, a resinous substance collected by honeybees called propolis, have been widely used in folk medicine since ancient times. Antibacterial, antifungal and thus antiseptic properties may represent the basis for the historical and present use of these extracts in dermatology, against inflammatory conditions and common colds. This work was carried out in order to verify possible biochemical alterations in some seric parameters of propolis-treated rats. It was shown that propolis possesses an antioxidant property and its administration did not affect either amylase and alanine transaminase activities or total protein concentration.

KEY WORDS: propolis; serum; biochemistry.

INTRODUCTION

Propolis is a resinous hive product, which has a versatile biological activity (7) . Bees use it to seal the hive walls and strengthen the borders of the combs as well as the hive entrance. In particular, they embalm dead invaders and other particles within the hive to prevent putrescence and microbial decomposition.

Propolis is in no way a new discovery. It has been in use since ancient times from where it found its way into the medicaments in folk medicine in many parts of the world. The ancient Egyptians used it to embalm their dead. In the Balkan States, propolis is still one of the most frequently used medication nowadays.

More than 180 propolis constituents have been identified by gas chromatography-mass spectrometry (GC-MS). These compounds can be grouped as follows: free aromatic acids; flavonoids; benzyl, methylbutenyl, phenylethyl, cinnamyl and other esters of these acids; chalcones and dihydrochalcones; terpenoids and others such as sugars, ketones, alcohols (2,8). Although in small quantities, these compounds can be very important to propolis activity (1). The identified constituents of propolis were mentioned by several authors (17).

Propolis has versatile biological activities, such as antibacterial (10), antiviral (14), immunostimulating (5), anti-inflammatory, among others. It is thought to be an antitumor agent, and it possibly presents a differentiation inducing agent (9), antioxidant properties (15), as well successful clinical applications, which has brought a greater interest in propolis effects.

In this work, we tried to verify whether propolis induces alterations in some biochemical parameters after its administration, and also to observe its antioxidant property. So, total protein, alanine aminotransferase (ALT - E.C. 2.6.1.2.), amylase (E.C. 3.2.1.1.) and Cu-Zn superoxide dismutase (SOD - E.C. 1.15.1.1.) determinations of the serum of propolis-treated rats were carried out.

MATERIAL AND METHODS

ANIMALS: Male Wistar rats weighing approximately 200g were used. The rats were acclimatized for one week, and received distilled water and Purina rat chow ad libitum throughout this study.

PROPOLIS: Propolis was obtained from the apiary located on Edgardia Farm (FMVZ - UNESP). A 30% propolis ethanolic solution was prepared and after a week this solution was filtered and used to prepare a 10% propolis hydroalcoholic solution (7).

TREATMENT OF THE ANIMALS: The animals were divided into three groups of five rats each. The control group (G1) received 0.7 ml of physiological saline solution (0.9 % NaCl - MERCK), four times a day. The other group (G2) received 0.7 ml of 10% hydroalcoholic solution (Labsynth) also four times a day, and the experimental group (G3) received four doses of 0.7 ml of 10% propolis hydroalcoholic solution. All the groups received the solutions through gavage during 24 hours.

SERUM SAMPLES: Twenty-four hours after propolis administration, rats were sacrificed by overetherization and decapitation. Blood samples were collected and centrifuged at 3,000 rpm for 10 minutes. The supernatant was used for total protein (11), ALT (18), amylase (3) and Cu-Zn superoxide dismutase (13) determinations.

STATISTICAL ANALYSIS: Analysis of variance was used to examine the effects of treatment and comparison between the means was performed by Tukey test. The probability of 0.05 was chosen as the significant level (19).

RESULTS

Propolis administration to rats showed no alterations in the seric levels of total protein (Table 1).

TABLE 1 -
Total protein, alanine aminotransferase (ALT), amylase and Cu-Zn superoxide dismutase activity (SOD) determination in the serum of control (G1), hydroalcoholic solution treated-rats (G2) and of propolis-treated rats (G3) 24h after treatment. Results are reported as means ± SD.

BIOCHEMICAL DETERMINATION	G1	G2	G3
Total protein (g/dL)	11.57 ± 1.84	10.85 ± 1.06	12.67 ± 1.00
SOD (U/mg protein)	16.71 ± 1.53	23.46 ±* 3.01	13.28 ± 4.33
ALT (U/L)	33.60 ± 0.65	31.77 ± 0.55	32.46 ± 2.30
Amylase (U/dL)	152.19 ± 2.14	151.83 ± 2.56	151.84 ± 1.68

Significantly increased serum Cu-Zn superoxide dismutase (SOD) activity was observed only in rats treated with hydroalcoholic solution, when compared to those of the control group and also to those treated with propolis. SOD activity was lower in the group which received propolis hydroalcoholic solution (Table 1; Figure 1).

FIGURE 1 -
Cu-Zn superoxide dismutase activity (SOD) of control, hydroalcoholic solution-treated rats and propolis-treated rats. Statistical analysis: alcohol > (propolis = control); p < 0.05.

There were no differences either in ALT or in amylase activities among the three groups (Table 1).

Table 1 summarizes all biochemical determinations; in general, no alterations were observed in biochemical parameters in the serum of propolis-treated rats.

DISCUSSION

Propolis has been reported to exhibit a wide range of properties, such as antibiotic, antiviral, anti-inflammatory, immunostimulatory, tumor carcinostatic, antioxidant, among others.

The purpose of this study was to investigate possible alterations in some biochemical parameters to observe whether propolis might lead to some damage in the tissues 24h after its administration.

There were no changes in the seric level of total protein in any group (Table 1). ALT and amylase showed no alterations in their activities either in the group treated with hydroalcoholic solution or in the group treated with propolis (Table 1). Since these enzymes are related to damage in the liver and in the pancreas, respectively, it might be hypothesized that propolis did not affect these tissues, and maybe the hydroalcoholic solution was weak to induce some alteration in these organs after 24h. Alterations in the seric levels of such enzymes have been thought to be a tool to study both cell viability and changes in cell membrane permeability.

On the other hand, it appeared that the hydroalcoholic solution induced superoxide generation, which induced SOD increased activity (Table 1; Figure 1). This enzyme shows an antioxidant activity, protecting highly unsaturated fatty acids in the lipids of biological membranes against deleterious effects of superoxide radical. Ethanol induces free radical generation, which is generally produced during the metabolism of aerobic living cells (12). The superoxide anion is an extremely potent free radical and because it is unstable, one can expect that even its low concentrations would be intolerable to the cells.

The ability to respond rapidly with increased antioxidant activity has been shown to be associated with relative tolerance to superoxide induced injury (6). Some authors determined the activity of SOD and lipid peroxidation and observed that the damage of liver mitochondrial membranes, shown in manganese deficient rats, may be due to depressed SOD activity resulting in an increased lipid peroxidation from free radicals (4).

Caffeic acid phenethyl ester, an active component of propolis extract, shows antioxidant properties by blocking the production of reactive oxygen species in human neutrophils (15). Water-soluble extracts show higher antioxidant activity when compared to the ethanolic extract. This demonstrates the medical superiority of propolis water-soluble extract due to the lack of allergic reactions, indicated in some cases for the ethanolic extract and the possibility of an oral administration especially in pediatrics or in ethanol-sensitive patients (16).

In this work we could verify that hydroalcoholic solution induced SOD increased activity, but the animals treated with propolis four times a day showed SOD normal activity. Once SOD activity was similar to that of the control group and the propolis-treated group, we might conclude that propolis exhibited an antioxidant property, in spite of being in a hydroalcoholic solution.

ACKNOWLEDGEMENTS

This study was performed with the technical assistance of Guerino Santo Bianchi Filho, Maria Conceição Tenore do Carmo and Luiz Carlos Fioravante. We thank Dr. Paulo Roberto Curi for the statistical analysis and Ana Claudia Acerra for the computer assistance.

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CORRESPONDENCE TO:

S.R.C. FUNARI, Departamento de Produção e Exploração Animal - FMVZ, UNESP - CEP 18.618-000 - Botucatu - São Paulo - Brasil.

Publication Dates

• Publication in this collection
  11 Jan 1999
• Date of issue
  1995